Product Name | Biotin - ACTH (1 - 39), humanBiotin - SYSMEHFRWGKPVGKKRRPVKVYPNGAEDESAEAFPLEF |
Size | 0.5 mg |
Catalog # | AS-23968 |
US$ | $232 |
Purity | % Peak Area By HPLC ≥ 95% |
This C-terminally labeled biotin ACTH (1-39) has been used in ELISA assays. Adrenocorticotropic hormone (ACTH), also known as corticotropin, is a cleavage product from a larger precursor proopiomelanocortin (POMC). This 39 amino acid-peptide hormone is produced in the anterior pituitary gland upon stimulation by the corticotropin releasing hormone from the hypothalamus in response to stress. It stimulates the secretion of steroid hormone, specifically glucocorticoids in the adrenal cortex by acting through a cell membrane receptor (ACTH-R). In mammals, the action of ACTH is limited to those areas of the adrenal cortex in which the glucocorticoid hormones cortisol (hydrocortisone) and corticosterone are formed. ACTH has little control over the secretion of aldosterone, the other major steroid hormone from the adrenal cortex. | |
Detailed Information | Material Safety Data Sheets (MSDS) |
Storage | -20°C |
References | Moreno-Guzmán, M. et al. Biosensors Bioelectronics 35, 82 (2012). doi: 10.1016/j.bios.2012.02.015Stewart, PM. et al. Clin Endocrinol 40, 199 (1994)Elias, LL. and AJ. Clark, Braz J Med Biol Res 33, 1245 (2000)Latronico, AC. Braz J Med Biol Res 33, 1249 (2000) |
Molecular Weight | 4767.6 |
Biotin-SYSMEHFRWGKPVGKKRRPVKVYPNGAEDESAEAFPLEF | |
Sequence(Three-Letter Code) | Biotin - Ser - Tyr - Ser - Met - Glu - His - Phe - Arg - Trp - Gly - Lys - Pro - Val - Gly - Lys - Lys - Arg - Arg - Pro - Val - Lys - Val - Tyr - Pro - Asn - Gly - Ala - Glu - Asp - Glu - Ser - Ala - Glu - Ala - Phe - Pro - Leu - Glu - Phe - OH |
Product Citations | Moreno-Guzmán, M. et al. (2012). Ultrasensitive detection of adrenocorticotropin hormone (ACTH) using disposable phenylboronic-modified electrochemical immunosensors. Biosensors Bioelectronics 35, 82. doi: 10.1016/j.bios.2012.02.015. |
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平常都是用超声破清洗器水浴超声破碎大约20分钟,然后上样的
但是一方面,常温超声不知道会不会造成蛋白降解破坏,因为发现加冰以后超声会变得非常弱,所以都么加冰。另一方面,因为水浴超声声强是不均匀的,如果样品多的话,会发现有些已经超碎DNA,但有些还是一团。
不知道是否有其他更好的方法?
1、安装电泳槽
将有机玻璃的电泳凝胶床洗净,晾干,用胶带将两端的开口封好,放在水平的工作台上,插上样品梳。
2、琼脂糖凝胶的制备
称取琼脂糖溶解在电泳缓冲液中,(按0.3-1.5%的琼脂糖含量,1-25kb大小的DNA用1%的凝胶,20-100kb的DNA用0.5%的凝胶,200-2000bp的DNA用1.5%的凝胶)置微波炉或沸水浴中加热至完全溶化(不要加热至沸腾),取出摇匀。
3、灌胶
将冷却到60℃的琼脂糖溶液轻轻倒入电泳槽水平板上。
4、待琼脂糖胶凝固后,在电泳槽内加入电泳缓冲液,然后拔出梳子。
5、加样
将DNA样品(DNA样品是细胞破碎之后用离心管离心沉淀获得的)与加样缓冲液按4:1混匀后,用微量移液器将混合液加到样品槽中,每槽加10-20μl,记录样品的点样次序和加样量。
6、电泳
安装好电极导线,点样孔一端接负极,另一端接正极,打开电源,调电压至3-5V/cm,电泳1-3hr,当溴酚蓝移到距凝胶前沿1-2cm时,停止电泳。
7、染色和观察
取出凝胶,放在含有溴化乙锭的染色液中染色30min,即可在254nm的紫外灯下观察,有橙红色荧光条带的位置,即为DNA条带,或在紫外灯下照相记录电泳图谱。溴化乙锭是致癌剂,操作时要小心,必须戴手套。
我怀疑溶菌酶有问题,因为其消化的30min内菌液几乎没大变化,而我曾看到别人用溶菌酶消化时菌液变的挺粘稠。
今下午再做时还是没什么变化,但发现溶菌酶在冰上静置2-3h后竟然沉淀下来了,而且上清和下面的白色溶菌酶分界明显,是我的溶菌酶配的有问题吗?出现这种情况溶菌酶还能用吗?我是-20度保存的。
下一步怎么处理菌液呢?
请高手多多指教!
暂无品牌问答